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Repair of a Critical-sized Calvarial Defect Model Using Adipose-derived Stromal Cells Harvested from Lipoaspirate
Published on: October 31, 2012
Peritoneal Submesothelial Stromal Cells Support Hematopoiesis and Differentiate into Osteogenic and Adipogenic Cell
R J F C do Amaral1, P Benac, L R Andrade
1Laboratório de Proliferação e Diferenciação Celular, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
Abstract:
The peritoneum is a thin membrane that covers most of the abdominal organs, composed of a monolayer of mesothelial cells and subjacent submesothelial loose connective tissue. Cells from the peritoneal wall are correlated with peritoneal fibrosis and epithelial-to-mesenchymal transition. However, the distinct involvement of mesothelial or submesothelial cells in such phenomena is still not clear. Here, we propose a new strategy to obtain stromal cells from anterior peritoneal wall explant cultures. These cells migrated from peritoneal tissues and proliferated in vitro for 4 weeks as adherent fibroblast-like cells. Optical and electronic microscopy analyses of the fragments revealed a significant submesothelial disorganization. The obtained cells were characterized as cytokeratin- vimentin+ laminin+ α-smooth muscle actin+, suggesting a connective tissue origin. Moreover, at the third passage, these stromal cells were CD90+CD73+CD29+Flk-1+CD45-, a phenotype normally attributed to cells of mesenchymal origin. These cells were able to support hematopoiesis, expressing genes involved in myelopoiesis (SCF, G-CSF, GM-CSF, IL-7 and CXCL-12), and differentiated into osteogenic and adipogenic cell lineages. The methodology demonstrated in this work can be considered an excellent experimental model to understand the physiology of the peritoneal wall in healthy and pathological processes. Moreover, this work shows for the first time that submesothelial stromal cells have properties similar to those of mesenchymal cells from other origins.
Insights
Researchers developed a new method to isolate peritoneal stromal cells, revealing their mesenchymal stem cell-like properties. These cells can support hematopoiesis and differentiate, offering a model for peritoneal wall research.
Area of Science:
- Cell Biology
- Tissue Engineering
- Connective Tissue Research
Background:
- The peritoneum, a mesothelial cell layer with subjacent connective tissue, is implicated in fibrosis and epithelial-to-mesenchymal transition.
- The specific roles of mesothelial versus submesothelial cells in these processes remain unclear.
Purpose of the Study:
- To develop a novel method for isolating and characterizing stromal cells from the anterior peritoneal wall.
- To investigate the properties and potential of these newly isolated peritoneal stromal cells.
Main Methods:
- Utilized explant cultures of the anterior peritoneal wall to obtain stromal cells.
- Employed light and electron microscopy to analyze tissue fragments.
- Characterized isolated cells using immunophenotyping (cytokeratin, vimentin, laminin, α-smooth muscle actin, CD90, CD73, CD29, Flk-1, CD45) and assessed their ability to support hematopoiesis and differentiate into osteogenic and adipogenic lineages.
Main Results:
- Successfully isolated and cultured fibroblast-like stromal cells from peritoneal explants.
- Microscopy revealed disorganization in the submesothelial layer.
- Characterization confirmed a connective tissue origin and a mesenchymal stem cell-like phenotype (CD90+CD73+CD29+Flk-1+CD45-).
- These cells supported hematopoiesis and demonstrated multi-lineage differentiation potential (osteogenic, adipogenic).
Conclusions:
- The developed methodology provides a valuable experimental model for studying peritoneal wall physiology in health and disease.
- This study demonstrates for the first time that submesothelial stromal cells possess mesenchymal stem cell-like properties, similar to those found in other anatomical locations.
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